The generation of TgMORN1 knockout (ΔTgMORN1) parasites.
A. Diagram describing the procedure for generating the ΔTgMORN1 parasite line. See text for details. B. Genomic PCR analysis of RHΔHX, the parental (LoxP-TgMORN1-HXGPRT-LoxP), and ΔTgMORN1 parasites. The diagram at the top shows the positions of the sequences that the PCR primers (A1, A2, S1, S2, S3) hybridize with. Boxes marked by slanted lines indicate regions of 5′ and 3′ UTR of TgMORN1 gene included in LoxP_TgMORN1_HXGPRT_LoxP knock-in plasmid (c.f. Figure 2A). C. Western blot analysis of RHΔHX, parental, ΔTgMORN1, and the complemented (ΔTgMORN1/eGFP-TgMORN1) parasites, which shows that the level of TgMORN1 expression was comparable among RHΔHX, the parental strain and the complemented parasites, but was undetectable in ΔTgMORN1 parasites. The blot was reprobed with mouse-anti-tubulin B-5-1-2 to use α-tubulin as a loading control.